Добавил:
Sekretar
kiopkiopkiop18@yandex.ru
t.me/Prokururor I Вовсе не секретарь, но почту проверяю
Опубликованный материал нарушает ваши авторские права? Сообщите нам.
Вуз:
Предмет:
Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5626_Библиотеки_им_академика_М_И_Перельмана.pdf
18.2 Plant Tissue Culture 361
Table 18.2 Examples of biotransformation reactions of plant cell and organ cultures.
Plant Precursor Product References
Astasia longa (R)- and (S)-Carvone Dihydrocarvone and
Catharanthus roseus (cell suspension
cultures)
Catharanthus roseus (cell suspension
cultures)
Centella asiatica Thiocolchicine 2-O- and 3-O-monoglucosyl
Daucus carota (immobilized plant cells) Codeinone Codeine [90]
Glycyrrhiza glabra (cell suspension
culture)
Peganum harmala (cell suspension
culture)
Rauwolfia serpentina (cell suspensions) Hydroquinone Arbutin [93]
certain significant secondary metabolite formation and
accumulation do not take place. Such cultures might still be
able to turn interesting products from external substrates,
though. The types of chemical substances that can go
through biotransformation mediated by plant enzymes are
varied [83]. The amount of enzyme activity present, the
presence of side reactions producing undesirable byproducts, the solubility of precursors, the localization of the
enzymes, and the presence of enzymes degrading the desired
product are just a few of the variables that will influence bioconversion rates by plant cells and organs. Cells’ ability to
convert nutrients into other forms can also be affected by
elicitation, permeabilization, pH changes, and osmotic
effects [84, 85]. Elicitors are biologically generated chemicals that induce secondary metabolite production; this stimulating process is known as elicitation. Endogenous elicitors
are elicitors generated within plant cells, such as pectin, pectic acid, cellulose, and other polysaccharides. Exogenous
elicitors, such as chitin, chitosan, and glucans are used to
describe elicitors that are produced by microbes. All biologically derived elicitors are biotic elicitors. Biotransformation
reactions performed by plant cell and organ cultures are
summarized in Table 18.2.
Vinblastine Vincristine [87]
Glychyrrhizin Glycyrrhetinic acid [88]
Papaverine Papaverinol [91]
Geranyl acetate, linalyl
acetate
3. Helpful in developing transgenic plants.
4. Tissue culture may be used by plant breeders to evalu-
ate cells rather than plants for advantageous features
such as herbicide tolerance or resistance.
5. Effective in haploid plant generation by anther or pol-
len culture.
6. Plant cells are cultivated in liquid culture in bioreac-
tors on a large scale to create valuable chemicals
including recombinant proteins for use as biopharmaceuticals and plant-derived secondary metabolites.
7. Biotransformation and secondary product biosynthesis
8. Useful for rapidly analyzing the molecular basis of
physiological, metabolic, and reproductive systems in
plants, such as stress-tolerant plant selection and flowering study in vitro.
9. To cause chromosomal doubling and polyploidy.
10. Meristem tip culture, as employed in fruit and potato
crops, can be utilized to grow virus-free plants from
virus-infected plants or stock.
11. The generation of sterile identical hybrid species can
be accomplished by tissue culture.
12. Biosynthetic pathways can be investigated with the
use of isolated organ and tissue cultures. An interpre-
isodihydrocarveol
derivatives
Geraniol, linalool,
alpha-terpineol
tation of the routes can be made by feeding the tissue
culture with the labeled precursor.
18.2.10 Applications of Plant Tissue Culture
Plant tissue culture has several applications which are
summarized below.
1. The production of several identical individuals from a
mother plant is possible through tissue culture.
2. To preserve endangered or rare plant species.
13. Mutant Selection: In terms of crop improvement, one
significant application of cell cultures is in mutant
selection. It is easier to isolate biochemical mutants
from cell culture than it is from the entire plant. It is
possible to screen a large number of cells for mutagenic therapies. Using this treatment cell lines resistant to fungal toxins, and herbicides are isolated.
[86]
[89]
[92]
362 18 Medicinal Plant Biotechnology
14. Creation of Artificial Seeds: Somatic embryos are
encapsulated in an appropriate matrix to create artificial or synthetic seeds. The apical and basal meristematic regions of somatic embryos have a bipolar
structure and can generate shoots and roots, respectively. Artificial seeds do not have endosperm or a seed
coat like zygotic embryos. In order to compensate for
these inadequacies, somatic embryos can be endospermized by encasing them in a suitable substance, such
as sodium alginate, and supplementing them with
growth regulators and nutrients. Longer storage times
for synthetic seeds do not result in a loss of viability.
Like regular seeds, they can be sown directly in the
ground.
15. Somaclonal Variations: These are genetic variations
with desired or improved characteristics that are introduced into plants in plant breeding programs to create
new varieties that can exhibit improved quality and
yield, disease resistance, and other traits in plants such
as cereals, legumes, oil seeds, tuber crops, etc. It is
easier to perform somaclonal variation than recombinant DNA technology. The Central Institute for
Medicinal and Aromatic Plants, Lucknow, India, has
released Bio-13, a medicinal plant that is a somaclonal
variety of Citronella java with 37% more oil, for commercial production.
18.3 Genetic Engineering (Recombinant DNA Technology)
In genetic engineering, a range of approaches are used to
purposefully modify the genetic material, typically deoxyribonucleic acid (DNA), of the host organism in order to
enhance its shape or function. Recombinant DNA techniques were developed in the second half of the twentieth century and frequently make use of bacteria or
bacteriophages as well as direct microinjection. Genetic
engineering is the systematic insertion of a foreign gene
or genes into the DNA of an organism. Heterologous
expression of foreign genes via suitable vectors can also
be achieved using genetic engineering. The genes can be
altered and reinserted into the same species, or they can
be separated and transferred from one species to another.
Transgenes, or new genes, are introduced into plants
through a process called transformation. The implanted
gene contains information that will provide the organism
with a new characteristic that it does not already have.
Recombinant DNA technology (rDNA) involves the following steps:
1. Generating DNA fragments or isolating genes of
interest,
2. DNA fragments are cut and joined to vector DNA
molecules,
3. Introducing vectors containing foreign DNA into host
cells so that they can multiply, and
4. Choosing the recipient cell clone(s) that have taken up
the recombinant DNA molecule.
There are several enzymes used in rDNA technology.
Enzymes, such as nucleases (Endonucleases, Exonucleases)
S1 nucleases, and DNases are used for cleaving (cutting)
DNA; DNA ligases are enzymes that are used to join DNA
fragments; DNA polymerase I, Terminal transferase, and
reverse transcriptase are used for amplification of DNA or
converting mRNA to DNA; alkaline phosphatase and
kinase are used to modify the ends of DNA molecule making them suitable for cloning; RNases are used to degrade
RNA. Methylases and calf intestinal phosphatase are some
other DNA-modifying enzymes used for the manipulation
of DNA fragments.
The separation of specific DNA fragments from the
entire genomic DNA is one of the most significant challenges for rDNA experiment. Normally, either DNA fragmentation or the synthesis of a new DNA molecule is used
for the experiment. Mechanical shearing is a technique for
fragmenting DNA molecules. Another advanced method
for generating DNA fragments is to use restriction endonucleases. Other methods for producing DNA fragments for
cloning include complementary DNA (cDNA) synthesis
utilizing mRNA as a template, followed by PCR-based
amplification of the target gene, i.e. cDNA.
18.3.1 Restriction Endonuclease
Restriction enzymes are bacterial enzymes that cut (cleave)
DNA at specified locations. Werner Arber revealed that
some enzymes defend the Escherichia coli (E. coli) bacterium from invading viral DNA by cutting and destroying it.
Restriction enzymes (REs) are enzymes that prevent viral
reproduction. REs recognize certain palindromic sequences
in double-stranded DNA that are four to six nucleotides
long and then cut both strands at specific locations. These
are known as recognition sequences. Type I REs are crucial
for bacterial function but do not break DNA at specific
points. Type II REs require very specific locations for DNA
breakage and are hence incredibly helpful tools in molecular biology. These enzymes make it possible to clone and
purify particular DNA segments. The approximately 500
known REs are frequently obtained from different bacterial strains. Most REs cleave recognition sequences on both
18.3 Genetic Engineering (Recombinant DNA Technology) 363
strands of DNA one or two base pairs distant from the
center. As a result, double-stranded DNA has short, singlestranded ends known as cohesive ends/sticky ends. Stickyend DNA fragments can easily link with other DNA pieces.
Adaptors can be used to connect the blunt ends. Adaptors
are short, chemically synthesized double strands of DNA
that may be used to connect the ends of two DNA molecules with differing sequences [94].
DNA Ligases: To reconnect the sliced DNA fragments,
DNA ligases are utilized. By catalyzing the formation of
a phosphodiester bond between the 3’ hydroxyl termini
of nucleotides and the 5’ phosphate group, DNA ligase
joins the DNA fragments [95]. The action of ligases is
not dependent on DNA sequence and will join bluntend termini as well as ends with cohesive overhanging
ends.
DNA Polymerases: All DNA Polymerases can catalyze
the addition of nucleotides to the 3’-OH ends of a primer
based in a template-directed manner and thus synthesizing the new DNA molecules. E. coli DNA Polymerase I,
T4 DNA polymerase, Taq DNA polymerase, reverse transcriptase, and other DNA polymerases have been characterized and are commercially accessible.
18.3.2 Vectors as Carriers of Transgene
A vector is a DNA molecule that carries foreign genetic
material into a different cell. A chimera is a vector that contains a foreign gene and is known as recombinant DNA. A
vector must contain (i) an ori site (Origin of replication),
(ii) multiple cloning site with RE sites and the ability to
insert foreign DNA into the vector, and (iii) the vector typically carries selectable markers, such as antibiotic resistance (e.g. tetracycline), allowing positive transformed cells
to be selected. Other desirable features that can be present
in a suitable cloning vector may be (i) vir genes for plant
transformation, (ii) lacZ fragment for complementation
and blue-white selection, (iii) integrase sites for chromosomal insertion, and (iv) to aid the purification of recombinant proteins after expression, reporter genes surround the
numerous cloning sites. Plasmids and bacteriophages are
the two most often employed forms of vectors.
A plasmid is a kind of extrachromosomal DNA that is
circular, double-stranded, and self-replicating. Plasmids
naturally give antibiotic resistance to the host bacterium.
They can take 6–10 kb fragments of DNA and reproduce
bacterial DNA independently. Plasmids are classified into
two types: conjugative and nonconjugative. Transfer genes
(tra) and mobilizing genes (mob) are found in conjugative
plasmids but not in nonconjugative plasmids. If the mob
gene is intact, nonconjugative plasmids can be mobilized
by another conjugative plasmid present in the same cell.
Bacteriophages, sometimes known as phages, are viruses
that attack and replicate inside bacteria. The unnecessary
phage genome which is one-third may be replaced by foreign DNA. These viral vectors may transport up to 23 kb of
DNA or RNA and contain viral promoters that allow the
target gene to be translated into the host cell. Phage λ and
phage M
are two regularly used phages. Vectors are fur-
13
ther subdivided into cloning and expression vectors based
on the purpose and stage of genetic engineering in which
they are utilized [96].
Categories of Vectors by Function
1. Cloning Vectors
The term “cloning vectors” refers to a class of vectors
that are used to replicate DNA fragments in an appropriate host. Due to the Ori (origin of replication site)
that a vector offers, it is utilized. Most cloning vectors
have synthetic multiple cloning sites, which contain
numerous restriction sites, in place of their natural
restriction sites to boost efficiency. Integrase sites (for
chromosomal insertion), vir genes (for plant transformation), and lacZa segment (for complementation)
are additional characteristics that can be added to vectors through engineering. A wide variety of cloning
vectors are available for microbial, plant and animal
genetic engineering applications.
2. Transcription Vectors
For any vector, transcription is a necessary element.
Stable transcription, which is dependent on vector promoters, is required for the stable expression of an
inserted gene. Vectors for transcription are only meant
to be duplicated or amplified, not translated or
expressed.
3. Expression Vectors
A vector is referred to be an expression vector when it
is created to express, or produce, the protein that the
DNA insert specifies. Vectors essentially have specific
promoter sequences and terminators to define the
expression cassette. Additionally, they may have features like His-Tag, GST-Tag, and reporter genes to facilitate the purification of recombinant proteins after
expression and production.
4. Shuttle Vectors
Shuttle vectors are plasmids that can spread genes
across two different species. As a result, they have two
replication origins, one for each host species, as well as
the replication genes that are not provided by the host
cells. The recombinant DNA technology is used to construct these vectors.
364 18 Medicinal Plant Biotechnology
18.3.3 Methods of Gene Transfer
18.3.3.1 Direct Gene Transfer Methods
When foreign DNA is inserted directly into the plant’s
genome, it is referred to as a “direct transfer of gene.” The
introduction of naked DNA into plant cells is the foundation of direct DNA transfer techniques.
Electroporation: Electric shocks can stimulate cellular
absorption of foreign DNA from a suspended solution
via holes created in the cell membrane by brief electric
pulses. It is a simple and rapid way of introducing genes
into the cells of numerous species. Plant protoplasts are
suspended using this approach in an appropriate ionic
solution containing linearized recombinant plasmid
DNA. After that, this combination is subjected to either
high-voltage short pulses or low-voltage long pulses for
the required number of cycles. It is believed that the
electrical pulses cause the plasma membrane to temporarily open holes, which allow the DNA molecules to be
integrated. Plants and cell colonies are then grown from
treated protoplasts. This process, which involves introducing DNA into plant cells by creating tiny holes in the
membrane, is known as electroporation.
Microprojectile Bombardment/Gene Gun
Sanford (1987) gave the microprojectile bombardment
technique its original name, biolistics. Ballistics and biology are combined to create biolistics. The target DNA is
covered in a carrier particle and then delivered into the tissue by firing the gene gun. This is the most general method
of transferring DNA into plant cells. The DNA of interest is
coated on to the tungsten or gold particles that range in
diameter from 1 to 5 m. The DNA-coated particles are
propelled into the target cell through a barrel at 430 m s
1
using compressed helium gas. Each DNA transfer event
requires about 50 g of tungsten. The carrier particles are
positioned in the particle acceleration device on a support
film. The support film is accelerated by gas pressure and
then halted by a protective mesh. The carrier particles contact the target tissue in a petri dish below the biolistic as
they travel through the mesh. The carrier particles enter
the mesh and make contact with the target. Below the
biolistic device is tissue mounted in a petri dish. When
there is a modest penetration number of bullets (1–5 per
cell), the target cell has a high chance of surviving. The
effectiveness of a gene gun depends on several variables,
including the chamber vacuum level, the range of particle
sizes, and the shot distance. The quantity of DNA per particle, the kind of explant, the physiological circumstances,
the type of gas, and the pressure are other important considerations. Important crop plants such as maize, rice, and
wheat have now been modified using this technique.
Microinjection
The desired DNA is mechanically inserted into a target cell
by a direct physical technique called microinjection. The
target cell could have been identified from meristems, callus, intact cells, protoplasts, embryos, etc. Microinjection is
a technique used to manipulate chromosomes and transfer
cellular organelles.
Microinjection is the act of inserting a cannula under a
microscope and introducing DNA straight into the cell or
even into the cell nucleus. Two pipette-equipped micromanipulators and a microcapillary needle (0.5–5 m in diameter) are used to hold the target cell in place. The injected
DNA is subsequently integrated into the plant’s genome
using the plant’s own DNA repair mechanism. The key
benefit is that no marker gene is required to identify successful transformation.
The recipient cells are kept immobile in agarose embedding and supported by a suction-holding pipette while the
gene transfer is being performed. After the microinjection
procedure is finished, the altered cell is cultivated and cultured to become a transgenic plant. This method has been
used to create transgenic tobacco and Brassica napus. The
main drawbacks of microinjection are that it requires specialized training, is costly, and takes long time.
Transformation: The process of introducing foreign DNA
into competent bacterial cells is known as transformation. The uptake of plasmid DNA by E. coli is carried out
in the presence of 0 to 5 °C ice-cold CaCl
quent heat shock (37–45 °C for 90 seconds). CaCl
and a subse-
2
alters
2
the permeability of the bacterial cell membrane and
makes the cell more permeable to take up exogenous
DNA. The transformational efficiency, or the proportion
of transformed cells, is high (one cell per 1000 cells).
Conjugation: During conjugation, single-stranded DNA is
transferred from the donor to the recipient cell by means
of cytoplasmic bridges formed when two living bacteria
(a donor and a recipient) unite.
Silicon Carbide Method: This technique transfers genes
using organic fibers, such as silicon carbide. These fibers
aid in the entry of foreign DNA into plant tissue when
combined with plasmid DNA and plant tissue or cells.
18.3.3.2 Indirect Gene Transfer Methods
Agrobacterium-mediated Gene Transfer Bacteria are used as a
vector in the indirect way of genetic transformation to
transfer the gene construct into the target cell. Agrobacterium
is used in this procedure. Agrobacterium tumefaciens
bacteria are home to the Ti plasmid (tumor-inducing),
which consists of transfer DNA (T-DNA) and other genes
required for T-DNA integration into the host genome.
Plants that have been injured generate sap that is rich in
18.3 Genetic Engineering (Recombinant DNA Technology) 365
phenolic chemicals, which act as chemical attractants for
agrobacteria and boost the expression of vir genes. It causes
Agrobacterium infection of the plant, insertion of the
T-DNA region at an undetermined location in the host
genome, and plant cell proliferation, which leads to crown
gall development. Agrobacterium rhizogens, which causes
plants to develop hairy roots, is a different species that is
frequently utilized. It possesses a root-inducing plasmid,
also known as the Ri plasmid. Numerous dicots and some
monocots are susceptible to infection by the genus
Agrobacterium, which has a broad range of hosts [97–100].
Structure of Ti Plasmid: The Ti plasmid comprises genes
for T-DNA integration, tumor induction, and the production of plant hormones and opines.
Origin of Replication: This region is in charge of Ti plas-
mid replication independent of the bacterial cell.
Virulence Region: This area comprises vir genes, the
products of which facilitate the processing and transmission of T-DNA from bacteria to plant cells. Plants emit
phenolic compounds like acetosyringone in reaction to
damage, which in turn induces their expression.
T-DNA Region: It is a section of the Ti plasmid that con-
tains genes from tumor induction. On both sides, it is
bordered by 25 bp direct repeat sequences. These repetitions are referred to as the Left border (LB) and Right
border (RB). This area contains the genes iaaM and
iaaH, which are responsible for the synthesis of indole
acetic acid (an auxin), ipt, which is responsible for the
manufacture of an enzyme isopentenyl adenine (a cytokinin), and tml, which is another gene implicated in
tumor development. Opine biosynthesis genes are
responsible for the production of opines. All of these
genes cause tissue expansion in plant cells, which leads
to cancer development.
Region of Opine Catabolism: It also comprises numer-
ous additional genes involved in opiate metabolism.
During infection, this portion of the plasmid is not transmitted to plant cells.
Use of Ti Plasmid in Genetic Transformation: For use
as a vector in genetic transformation, the majority of the
T-DNA region of the bacterial plasmid is replaced with
the gene of interest, but the left and right border
sequences are left alone. The T-DNA region is specified
by its boundaries rather than its sequence, which allows
it to be inserted into the host plant genome.
18.3.4 Applications of Genetic Engineering
Agriculture: The most well-known use of genetic engineer-
ing is in agriculture, where researchers have employed
modern technology to improve selective breeding in order
to improve the qualities of future crop generations. Classic
examples include the development of insect-resistant cotton crops, drought-resistant varieties of crop plants, and
“biofortified” food crops with improved nutritional value
like Golden Rice and Multivitamin Corn. Biofortified
foods, such as Golden Rice, that contain genes for
increased β-carotene content, a precursor of vitamin A,
have played an essential role in addressing vitamin A
insufficiency in developing countries. Over the years, several biofortified varieties of pulses, cereals, vegetables, oilseeds, and fruits have been developed. Second-generation
sophisticated gene editing technologies, such as CRISPR/
Cas9, which permits precise genome changes, have been
widely employed over the last decade to modify model
plant genomes as well as crop species for yield enhancement and biotic and abiotic stress management. Metabolic
engineering of plants is yet another facet to modify endogenous pathways of plants in order to increase the production of desired metabolites such as alkaloids, terpenoids,
or specific valuable metabolites which can be of use in
medicine, industry, or increase the innate plant defense.
Metabolic engineering of volatile organic compounds can
significantly improve the natural plant defense and serve
as an alternate pest management strategy.
Environmental Management: For bioremediation, also
known as phytoremediation, genetically engineered
plants have been developed. A method for removing or
neutralizing environmental toxins, heavy metals and
metalloids is known as phytoremediation. It can be
accomplished by either breeding or engineering plants
with improved or novel capabilities or by utilizing the
inherent properties of plants in environmental management regimes. Genetic engineering can be employed to
strengthen the phytoremediation capacity of plants.
Plants, for example might be genetically engineered to
behave as heavy metal magnets in soil and water or to
generate more enzymes that can biodegrade materials
and harmful contaminants. Effective approaches include
exploiting the genes involved in metal uptake, translocation, reduction, and vacuolar sequestration. Genetically
modified versions of plants can be designed to enhance
their ability to reduce indoor air pollution.
Biopharmaceuticals: Genetic engineering, for instance,
is helpful in drug discovery for the synthesis of novel
therapeutic agents as well as the invention and improvement of novel techniques to consistently generate those
molecules. Plants can be genetically engineered to generate pharmacologically active proteins such as antibodies, vaccines, hormones, cytokines, and a range of
medicinal medicines. This domain is popularly known
as “Biopharming” as it turns plants into biofactories to
produce inexpensive and ingestible medicines.
366 18 Medicinal Plant Biotechnology
18.4 Conclusion
Finding new products of medicinal importance from plants
is a very prominent area of research and a valuable resource
that further needs much exploration. Many plants, including wild species, have recently been identified and validated
as valuable sources of natural chemicals for pharmacy and
medicine. Recent advances in molecular biology and genetic
engineering of plant cell cultures suggest that these systems
can be transformed into substantial secondary metabolite
sources. The transgenic plants can be of significant use to
produce protein at an uninterrupted pace. In-vitro propagation is a highly advanced and commercialized field worldwide from an applied perspective. Every year, a huge number
of laboratories create several plants, mostly those that are
vegetatively propagated, including flowers, grapes, ornamentals, fruit trees, and rootstocks. Furthermore, the emergence of advanced gene editing technologies has
revolutionized the field of medicinal plant biotechnology.
Developing sustainable agriculture technologies is crucial
for ensuring food security and developing economies.
References
1 Dias, D.A., Urban, S. and Roessner, U. (2012). A
historical overview of natural products in drug
discovery. Metabolites 2 (2): 303–36. https://doi.org/10.
3390/metabo2020303.
2 Bandaranayake, W.M. (2006). Quality control, screening,
toxicity, and regulation of herbal drugs. In: Modern
Phytomedicine: Turning Medicinal Plants into Medicines
(ed. I. Ahmad, F. Aqil and M. Owais), 25–57.
Weinheim: Wiley-VCHGmbH & Co. KGaA. https://doi.
org/10.1002/9783527609987.ch2.
3 Tugume, P. and Nyakoojo, C. (2019). Ethno-
pharmacological survey of herbal remedies used in the
treatment of paediatric diseases in Buhunga parish,
Rukungiri district, Uganda. BMC Complementary and
Alternative Medicine 19 (1): 353. https://doi.org/10.1186/
s12906-019-2763-6.
4 Grafi, G. (2004). How cells dedifferentiate: a lesson from
plants. Developmental Biology 268 (1): 1–6. https://doi.org/
10.1016/j.ydbio.2003.12.027.
5 Guzzo, F., Baldan, B., Mariani, P. et al. (1994). Studies
on the origin of totipotent cells in explants of Daucus
carota L. Journal of Experimental Botany 45 (279):
1427–32. http://www.jstor.org/stable/23694642.
6 Gaillochet, C. and Lohmann, J.U. (2015). The never-
ending story: from pluripotency to plant developmental
plasticity. Development (Cambridge, England) 142 (13):
2237–49. https://doi.org/10.1242/dev.117614.
7 Thorpe T.A. (2007). History of plant tissue culture.
Molecular Biotechnology 37 (2): 169–80. https://doi.org/
10.1007/s12033-007-0031-3.
8 Gautheret, R.J. (1983). Plant tissue culture: A history.
Botanical Magazine, Tokyo 96: 393–10. https://doi.org/
10.1007/BF02488184.
9 Gautheret, R.J. (1955). The nutrition of plant tissue
cultures. Annual Review of Plant Physiology 6 (1):
433–84. https://doi.org/10.1146/annurev.pp.06.060155.
002245
10 De Fossard, R.A. (1986). Principles of plant tissue
culture. In: Tissue Culture as a Plant Production System
for Horticultural Crops. Current Plant Science and
Biotechnology in Agriculture (ed. R.H. Zimmerman, R.J.
Griesbach, F.A. Hammerschlag. et al.), 1–13. Dordrecht:
Springer. https://doi.org/10.1007/978-94-009-4444-2_1.
11 George, E.F., Hall, M.A. and Klerk, G.J.D. (2008). The
components of plant tissue culture media I: macro- and
micro-nutrients. In: Plant Propagation by Tissue Culture
(ed. E.F. George, M.A. Hall and GJ.D. Klerk), 65–113.
Dordrecht: Springer. https://doi.org/10.1007/978-1-4020
-5005-3_3.
12 Biedermann, I.E.G. (1987). Factors affecting
establishment and development of Magnolia hybrids in
vitro. ISHS Acta Horticulturae 212: 625–29. https://doi.
org/10.17660/ActaHortic.1987.212.104.
13 Chin, C. and Miller, D. (1982). Some characteristics of
the phosphate uptake by Petunia cells. HortScience
17 (3 Section 2): 488.
14 Pasqualetto, P.L., Zimmerman, R.H. and Fordham, I.
(1988). The influence of cation and gelling agent
concentrations on vitrification of apple cultivars in vitro.
Plant Cell Tissue and Organ Culture 14: 31–40. https://
doi.org/10.1007/BF00029573.
15 White, P.J. and Broadley, M.R. (2003). Calcium in plants.
Annals of Botany 92 (4): 487–11. https://doi.org/
10.1093/aob/mcg164.
16 Khan, F., Siddique, A.B., Shabala, S. et al. (2023).
Phosphorus plays key roles in regulating plants’
Switzerland) 12 (15): 2861. https://doi.org/10.3390/plant
s12152861.
17 Ishfaq, M., Wang, Y., Yan, M. et al. (2022). Physiological
essence of magnesium in plants and its widespread
deficiency in the farming system of China. Frontiers in
Plant Science 13: 802274. https://doi.org/10.3389/fpls.
2022.802274.
18 Narayan, O.P., Kumar, P., Yadav, B. et al. (2022). Sulfur
nutrition and its role in plant growth and development.
367References
Plant Signaling & Behavior 18 (1): 2030082. https://doi.
org/10.1080/15592324.2022.2030082.
19 Sommer, A.L. and Lipman, C.B. (1926). Evidence on
the indispensable nature of zinc and boron for higher
green plants. Plant Physiology 1 (3): 231–49. https://
doi.org/10.1104/pp.1.3.231.
20 Sommer, A.L. (1931). Copper as an essential for plant
growth. Plant Physiology 6 (2): 339–45. https://doi.org/
10.1104/pp.6.2.339.
21 Arnon, D.I. and Stout, P.R. (1939). Molybdenum as an
essential for higher plants. Plant Physiology 14 (3):
599–2. https://doi.org/10.1104/pp.14.3.599.
22 Alejandro, S., Höller, S., Meier, B. et al. (2020).
Manganese in plants: from acquisition to subcellular
allocation. Frontiers in Plant Science 11: 300. https://
doi.org/10.3389/fpls.2020.00300.
23 Klein, R.M., Caputo, E.M. and Witterholt, B.A. (1962).
The role of zinc in the growth of plant tissue cultures.
American Journal of Botany 49 (4): 323–27. https://
doi.org/10.2307/2439069.
24 Sommer, A.L. and Lipman, C.B. (1926). Evidence on the
indispensable nature of zinc and boron for higher green
plants. Plant Physiology. 1: 231–49
25 Whittington, W.J. (1959). The role of boron in plant
growth: The effect on growth of the radicle. Journal of
Experimental Botany 10 (28): 93–3.
26 Shireen, F., Nawaz, M. A., Chen, C. et al. (2018). Boron:
functions and approaches to enhance its availability in
plants for sustainable agriculture. International Journal of
Molecular Sciences 19 (7): 1856. https://doi.org/10.3390/
ijms19071856.
27 Dahleen, L.S. (1995). Improved plant regeneration from
barley callus cultures by increased copper levels. Plant
Cell, Tissue and Organ Culture 43: 267–69. https://
doi.org/10.1007/BF00039954.
28 Tamimi, S.M. and Othman, H. (2020). Effects of copper
sulphate on shoot multiplication and rooting of banana
(Musa acuminata L.) (In vitro Study). Asian Journal of
Plant Sciences 19: 200–4. https://doi.org/10.3923/ajps.
2020.200.204.
29 Hu, X., Wei, X., Ling, J. et al. (2021). Cobalt: an essential
micronutrient for plant growth? Frontiers in Plant
Science 12: 768523. https://doi.org/10.3389/fpls.2021.
768523.
30 Colmenero-Flores, J.M., Franco-Navarro, J.D., Cubero-
Font, P. et al. (2019). Chloride as a beneficial
macronutrient in higher plants: new roles and
regulation. International Journal of Molecular Sciences
20 (19): 4686. https://doi.org/10.3390/ijms20194686.
31 Broyer, T.C., Carlton, A.B., Johnson, C.M. et al. (1954).
Chlorine-A micronutrient element for higher plants.
Plant Physiology 29 (6): 526–32. https://doi.org/10.1104/
pp.29.6.526.
32 Johnson, C.M., Stout, P.R. and Broyer, T.C. (1957).
Comparative chlorine requirements of different plant
species. Plant Soil 8: 337–53. https://doi.org/10.1007/
BF01666323.
33 Kaiser, B.N., Gridley, K.L., Ngaire Brady, J. et al. (2005).
The role of molybdenum in agricultural plant
production. Annals of Botany 96 (5): 745–54. https://
doi.org/10.1093/aob/mci226.
34 Rout, G.R. and Sahoo, S. (2015). Role of iron in plant
growth and metabolism. Reviews in Agricultural Science
3: 1–24. https://doi.org/10.7831/ras.3.1.
35 Kolthoff, I.M. and Auerbach, C. (1952). Studies on the
system iron-ethylenediamine tetraacetate. Journal of
the American Chemical Society 74 (6): 1452–56. https://
doi.org/10.1021/ja01126a027.
36 Legrand, B. (1975). Action of iron and EDTA on the
neoformation of buds, by the leaf fragments of endive
cultivated in vitro. Comptes Rendus De l’Académie Des
Sciences 280D: 2215–18.
37 White P.R. (1938). Accessory salts in the nutrition of
excised tomato roots. Plant Physiology 13 (2): 391–98.
https://doi.org/10.1104/pp.13.2.391.
38 Kiferle, C., Martinelli, M., Salzano, A.M. et al. (2021).
Evidences for a nutritional role of iodine in plants.
Frontiers in Plant Science 12: 616868. https://doi.org/10.
3389/fpls.2021.616868.
39 Fabiano, C.C., Tezotto, T., Favarin, J.L. et al. (2015).
Essentiality of nickel in plants: a role in plant stresses.
Frontiers in Plant Science 6: 754. https://doi.org/10.3389/
fpls.2015.00754.
40 Sivanesan, I. and Park, S.W. (2014). The role of silicon in
plant tissue culture. Frontiers in Plant Science 5: 571.
https://doi.org/10.3389/fpls.2014.00571.
41 Yaseen, M., Ahmad, T., Sablok, G. et al. (2013). Review:
Role of carbon sources for in vitro plant growth and
development. Molecular Biology Reports 40 (4): 2837–49.
https://doi.org/10.1007/s11033-012-2299-z.
42 Leifert, C., Murphy, K.P. and Lumsden, P.J. (1995).
Mineral and carbohydrate nutrition of plant cell and
tissue cultures. Critical Reviews in Plant Sciences 14 (2):
83–109, https://doi.org/10.1080/07352689509701923.
43 Fitzpatrick, T.B. and Chapman, L.M. (2020). The
importance of thiamine (vitamin B
) in plant health:
1
From crop yield to biofortification. The Journal of
Biological Chemistry 295 (34): 12002–13. https://doi.
org/10.1074/jbc.REV120.010918.
44 Mohamed, G.M., Amer, A.M., Osman, N.H. et al. (2021).
Effects of different gelling agents on the different stages
of rice regeneration in two rice cultivars. Saudi Journal
368 18 Medicinal Plant Biotechnology
of Biological Sciences 28 (10): 5738–44. https://doi.org/
10.1016/j.sjbs.2021.06.003.
45 Gaspar, T., Kevers, C., Penel, C. et al. (1996). Plant
hormones and plant growth regulators in plant tissue
culture. In Vitro Cellular and Developmental Biology-
Plant 32: 272–89.
46 Hill, K. and Schaller, G.E. (2013). Enhancing plant
regeneration in tissue culture: a molecular approach
through manipulation of cytokinin sensitivity. Plant
Signaling & Behavior 8 (10): https://doi.org/10.4161/
psb.25709.
47 Grzegorczyk-Karolak, I., Hnatuszko-Konka, K.,
Krzemińska, M. et al. (2021). Cytokinin-based tissue
cultures for stable medicinal plant production:
Regeneration and phytochemical profiling of Salvia
bulleyana shoots. Biomolecules 11 (10): 1513. https://doi.
org/10.3390/biom11101513.
48 Castro-Camba, R., Sánchez, C., Vidal, N. et al. (2022).
Plant development and crop yield: the role of
gibberellins. Plants (Basel, Switzerland) 11 (19): 2650.
https://doi.org/10.3390/plants11192650.
49 Long, Y., Yang, Y., Pan, G. et al. (2022). New insights
into tissue culture plant-regeneration mechanisms.
Frontiers in Plant Science 13: 926752. https://doi.org/10.
3389/fpls.2022.926752.
50 Bhojwani, S.S. and Dantu, P.K. (2013).
Micropropagation. In: Plant Tissue Culture: An
Introductory Text, 245–74. India: Springer. https://doi.
org/10.1007/978-81-322-1026-9_17.
51 Etienne, H. and Berthouly, M. (2002). Temporary
immersion systems in plant micropropagation. Plant
Cell, Tissue and Organ Culture 69: 215–31. https://doi.
org/10.1023/A:1015668610465.
52 Gupta, N., Jain, V., Joseph, M. et al. (2020). A review on
micropropagation culture method. Asian Journal of
Pharmaceutical Research and Development 8 (1): 86–3.
https://doi.org/10.22270/ajprd.v8i1.653.
53 Hayta, S., Bayraktar, M., Baykanerel, S. et al. (2017).
Direct plant regeneration from different explants
through micropropagation and determination of
secondary metabolites in the critically endangered
endemic Rhaponticoides mykalea. Plant Biosystems
151 (1): 20–8. https://doi.org/10.1080/11263504.2015.
1057267.
54 Sidhu, Y. (2011). In vitro micropropagation of medicinal
plants by tissue culture. The Plymouth Student Scientist
4 (1): 432–49.
55 Guan, Y., Li, S.G., Fan, X.F. et al. (2016). Application of
somatic embryogenesis in woody plants. Frontiers in Plant
Science 7: 938. https://doi.org/10.3389/fpls.2016.00938.
56 Gleddie, S., Keller, W. and Setterfield, G. (1983).
Somatic embryogenesis and plant regeneration from
leaf explants and cell suspensions of Solanum
melongena (eggplant). Canadian Journal of Botany
61: 656–66.
57 Dodeman, V.L., Ducreux, G. and Kreis, M. (1997).
Zygotic embryogenesis versus somatic embryogenesis.
Journal of Experimental Botany 48: 1493–1509.
58 Jayasankar, S., Gray, D.J. and Litz, R.E. (1999). High-
efficiency somatic embryogenesis and plant
regeneration from suspension cultures of grapevine.
Plant Cell Report 18: 533–37
59 Park, Y.S., Barrett, J.D. and Bonga, J.M. (1998)
Application of somatic embryogenesis in high value
clonal forestry: Development, genetic control and
stability of cryopreserved clones. In Vitro Cellular &
Developmental Biology - Plant 34: 231–39
60 Trolinder N.L. and Goodin J.R. (1988) Somatic
embryogenesis in cotton (Gossypium). II. Requirements
for embryo development and plant regeneration. Plant
Cell Tissue Organ Culture 12: 43–53.
61 Miller, R.A., Kao, K.N. and Gamborg, O.L. (1973). Plant
protoplast culture11NRCC No. 13002. In: Tissue Culture
(ed. P.F. Kruse and M.K. Patterson), 500–505. London:
Academic Press. https://doi.org/10.1016/B978-0-12427150-0.50117-0.
62 Wiszniewska, A. and Pindel, A. (2010). Protoplast
culture utilization in studies on legume crops. Acta
Agriculturae Scandinavica, Section B-Soil & Plant
Science 60 (5): 389–99. https://doi.org/10.1080/
09064710903179545.
63 Espinosa-Leal, C.A., Puente-Garza, C.A. and García-
Lara, S. (2018). In vitro plant tissue culture: means for
production of biological active compounds. Planta
248 (1): 1–18. https://doi.org/10.1007/s00425-018-2910-1.
64 Chandran, H., Meena, M., Barupal, T. et al. (2020). Plant
tissue culture as a perpetual source for production of
industrially important bioactive compounds.
Biotechnology Reports (Amsterdam, Netherlands)
26: e00450. https://doi.org/10.1016/j.btre.2020.e00450.
65 Ara, H., Jaiswal, U. and Jaiswal, V.S. (2000). Synthetic
seed: Prospects and limitations. Current Science 78 (12):
1438–44.
66 Gray, D.J., Compton, M.E., Harrell, R.C. et al. (1995).
Somatic embryogenesis and the technology of synthetic
seed. In: Somatic Embryogenesis and Synthetic Seed I.
Biotechnology in Agriculture and Forestry (ed. Y.P.S.
Bajaj), Berlin: Springer. https://doi.org/10.1007/978-3-66
2-03091-2_9.
67 Helal, N.A.S. (2011). The green revolution via synthetic
(artificial) seeds: A review. Research Journal of
Agriculture and Biological Sciences 7 (6): 464–77.
68 Khor, E. and Loh, C.S. (2005). Artificial seeds. In:
Applications of Cell Immobilisation Biotechnology. Focus
369References
on Biotechnology (ed. T. Page), 527–537. Dordrecht:
Springer. https://doi.org/10.1007/1-4020-3363-X_31.
69 Kitto, S.K. and Janick, J. (1982). Polyox as an artificial
seed coat for asexual embryos. Hort Science 17: 488–90.
70 Onishi, N., Sakamoto, Y. and Hirosawa, T. (1994).
Synthetic seeds as an application of mass production of
somatic embryos. Plant Cell, Tissue and Organ Culture
39: 137145. https://doi.org/10.1007/BF00033921.
71 Nausch, H. and Buyel, J.F. (2021). Cryopreservation of
plant cell cultures - Diverse practices and protocols. New
Biotechnology 62: 86–5. https://doi.org/10.1016/j.nbt.
2021.02.002.
72 Bai, F.W., Zhao, X.Q. and Xu, J. (2011). Immobilization
technology: Cells. In: Comprehensive Biotechnology
(ed. M. Moo-Young), 477–489. London: Academic Press.
https://doi.org/10.1016/B978-0-08-088504-9.00115-X.
73 Karel, S.F., Libicki, S.B. and Robertson, C.R. (1985). The
immobilization of whole cells: Engineering principles.
Chemical Engineering Science 40 (8): 1321–54. https://
doi.org/10.1016/0009-2509(85)80074-9.
74 Akin, C. (1987). Biocatalysis with immobilized cells.
Biotechnology & Genetic Engineering Reviews 5: 319–67.
https://doi.org/10.1080/02648725.1987.10647842.
75 Dervakos, G.A. and Webb, C. (1991). On the merits of
viable-cell immobilisation. Biotechnology Advances 9 (4):
559–12. https://doi.org/10.1016/0734-9750(91)90733-c.
76 Groboillot, A., Boadi, D.K., Poncelet, D. et al. (1994).
Immobilization of cells for application in the food
industry. Critical Reviews in Biotechnology 14 (2):
75–107. https://doi.org/10.3109/07388559409086963.
77 Norton, S. and D’Amore, T. (1994). Physiological effects
of yeast cell immobilization: Applications for brewing.
Enzyme and Microbial Technology 16 (5): 365–75.
https://doi.org/10.1016/0141-0229(94)90150-3.
78 Park, J.K. and Chang, H.N. (2000). Microencapsulation
of microbial cells. Biotechnology Advances 18 (4):
303–19. https://doi.org/10.1016/s0734-9750(00)00040-9.
79 Rathore, S., Desai P.M., Liew C.V. et al. (2013).
Microencapsulation of microbial cells. Journal of Food
Engineering 116 (2): 369–81. https://doi.org/10.1016/j.
jfoodeng.2012.12.022.
80 Mavituna, F. (2004). Pre-formed carriers for cell
immobilisation. In: Fundamentals of Cell Immobilisation
Biotechnology. Focus on Biotechnology (ed. V. Nedović
and R. Willaert), 121–139. Dordrecht: Springer. https://
doi.org/10.1007/978-94-017-1638-3_7.
81 Lindsey, K. (1985). Manipulation, by nutrient limitation,
of the biosynthetic activity of immobilized cells of
Capsicum frutescens Mill. cv. annuum. Planta 165 (1):
126–33. https://doi.org/10.1007/BF00392221.
82 Ishida, B.K. (1988). Improved diosgenin production in
Dioscorea deltoidea cell cultures by immobilization in
polyurethane foam. Plant Cell Reports 7 (4): 270–73.
https://doi.org/10.1007/BF00272540.
83 Franssen, M.C.R. and Walton, M.J. (1999).
Biotransformations. In: Chemicals From Plants,
Perspectives on Plant Secondary Products (ed. N.J. Walton
and D.E. Brown), 277–325. London: Imperial College
Press. https://doi.org/10.1142/9789812817273_0007.
84 Tang, Y.X. and Suga, T. (1994). Biotransformation of
alpha- and beta-ionones by immobilized cells of
Nicotiana tabacum. Phytochemistry 37 (3): 737–40.
https://doi.org/10.1016/s0031-9422(00)90349-2.
85 Fowler, M.W. and Stafford, A.M. (1992). Plant cell
culture, process systems and product synthesis. In: Plant
Biotechnology (ed. M.W. Fowler and G.S. Warren),
79–98. Oxford: Pergamon.
86 Shimoda, K. and Hirata, T. (2000). Biotransformation of
enones with biocatalysts-two enone reductases from
Astasia longa. Journal of Molecular Catalysis B:
Enzymatic 8 (4–6): 255–64. https://doi.org/10.1016/
S1381-1177(99)00076-4.
87 Hamada, H. and Nakazawa, K. (1991).
Biotransformation of vinblastine to vincristine by cell
suspension cultures of Catharanthus roseus.
Biotechnology Letters 13: 805–6. https://doi.org/10.
1007/BF01026763.
88 Hamada, H. and Nakata, S. (1992). Biotransformation of
glycyrrhizin by buffered cell suspension cultures of
Catharanthus roseus. Plant Tissue Culture Letters 9: 32–3.
https://doi.org/10.5511/plantbiotechnology1984.9.32.
89 Solet J.M., Bister-Miel, F., Galons, H. et al. (1993).
Glucosylation of thiocolchicine by a cell suspension
culture of Centella asiatica. Phytochemistry 33 (4):
817–20. https://doi.org/10.1016/0031-9422(93)85281-U.
90 Jones, A. and Veliky, I.A. (1981). Biotransformation of
cardenolides by plant cell cultures1. Planta medica
42 (6): 160–66. https://doi.org/10.1055/s-2007-971620.
91 Dorisse, P., Gleye, J., Loiseau, P. et al. (1988). Papaverine
biotransformation in plant cell suspension cultures.
Journal of Natural Products 51 (3): 532–36. https://doi.
org/10.1021/np50057a013.
92 Zhu, W. and Lockwood, G. (2000). Enhanced
biotransformation of terpenes in plant cell suspensions
using controlled release polymer. Biotechnology Letters
22: 659–62. https://doi.org/10.1023/A:1005675417538.
93 Lutterbach, R. and Stöckigt, J. (1992). High-yield
formation of arbutin from hydroquinone by cellsuspension cultures of Rauwolfia serpentina. Helvetica
Chimica Acta 75: 2009–11. https://doi.org/10.1002/hlca.
19920750625.
94 Davis, L.G., Dibner, M.D. and Battey, J.F. (1986).
Restriction endonucleases (Res) and their use. In: Basic
Methods in Molecular Biology (ed. L.G. Davis, M.D. Dibner
370 18 Medicinal Plant Biotechnology
and J.F. Battey), 51–57. New York: Elsevier. https://
doi.org/10.1016/B978-0-444-01082-7.50021-7.
95 Tomkinson, A.E. and Della-Maria, J.A. (2013). DNA
ligases: mechanism and functions. In: Encyclopedia of
Biological Chemistry (ed. W.J. Lennarz and M.D. Lane),
28–32. London: Academic Press. https://doi.org/10.1016/
B978-0-12-378630-2.00303-0.
96 Zaidi, S.S. and Mansoor, S. (2017). Viral vectors for
plant genome engineering. Frontiers in Plant Science
8: 539. https://doi.org/10.3389/fpls.2017.00539.
97 Gelvin, S.B. (2003). Agrobacterium-mediated plant
transformation: The biology behind the “genejockeying” tool. Microbiology and Molecular Biology
Reviews: MMBR 67 (1): 16–37. https://doi.org/10.1128/
MMBR.67.1.16-37.2003.
98 Abuodeh, R.O., Orbach, M.J., Mandel, M.A. et al. (2000).
Genetic transformation of Coccidioides immitis
facilitated by Agrobacterium tumefaciens. The Journal of
Infectious Diseases 181 (6): 2106–10. https://doi.org/10.
1086/315525.
99 Allen, G.C., Hall, G.E., Childs, L.C. et al. (1993). Scaffold
attachment regions increase reporter gene expression in
stably transformed plant cells. The Plant Cell 5 (6):
603–13. https://doi.org/10.1105/tpc.5.6.603.
100 Anandalakshmi, R., Pruss, G.J., Ge, X. et al. (1998). A
viral suppressor of gene silencing in plants. Proceedings of
the National Academy of Sciences of the United States of
America 95 (22): 13079–84. https://doi.org/10.1073/
pnas.95.22.13079.